LOCUS       sequence-8722-ex        2984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N-term GST fusion, frame 6P-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2984)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 2984)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pENTR4-GST 6P-1 (w487-1)
FEATURES             Location/Qualifiers
     source          1..2984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             475..1128
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1086..1108
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1135..1158
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    complement(1214..1313)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1425..1444)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1436..2245
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1503..1522)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2335..2923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2824..2843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggtaga tctcatgtcc
      481 cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa
      541 tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga
      601 aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat
      661 gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg
      721 ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt
      781 agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt
      841 cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat
      901 ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt
      961 ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt
     1021 attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct
     1081 ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctggaa
     1141 gttctgttcc aggggcccct gggatccccg gaattcccgg gtcgactcga gcggccgcac
     1201 tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1261 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1321 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1381 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1441 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1501 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1561 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1621 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1681 caagcatttt atccgtactc ctggtgatgc atggttactc accactgcga tccccggaaa
     1741 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     1801 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     1861 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     1921 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     1981 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2041 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2101 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2161 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2221 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac attattcaga
     2281 ttgggccccg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2461 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
